Fig 2: CASC11 positively regulated RAB11FIP2 expression in CRC cells. (A-D) The relative expression levels of RAB11FIP2 in CRC cells transfected with CASC11 overexpression or interference reagents were determined by qRT-PCR and WB. (E) Expression of CASC11 and RAB11FIP2 in CRC tissues and the relationship between them (n=27). Left panel, qRT-PCR analysis of CASC11 and RAB11FIP2 expression in 27 paired human CRC tissues and adjacent normal tissues. Right panel, spearman correlation analysis showed a positive relationship between CASC11 expression levels and RAB11FIP2 mRNA levels in 27 CRC tissue samples. Error bars represent the means ± SD of 3 independent experiments. **p < 0.01; ***p <0.001.

Fig 3: miR-646 and miR-381-3p co-target and negatively regulate RAB11FIP2. (A) Predicted binding sites of miR-646 and miR-381-3p on the RAB11FIP2 3’UTR sequence. The white nucleotides are the seed sequences of miR-646 and miR-381-3p. (B, C) Luciferase activities were measured in CRC cells co-transfected with a luciferase reporter containing the wild-type RAB11FIP2 3’UTR and miR-646 and miR-381-3p mimics or the mutant RAB11FIP2 3’UTR. Data are presented as the relative ratio of Renilla luciferase activity and firefly luciferase activity. (D) The relative expression levels of RAB11FIP2 in CRC cells transfected with miR-646 and miR-381-3p mimics or inhibitors were determined by qRT-PCR and WB. Error bars represent the means ± SD of 3 independent experiments. ns, no significance; **p < 0.01; ***p <0.001.

Fig 4: CASC11 is a PI3K/AKT signalling pathway regulator in CRC cells. (A) The levels of the main molecules of the PI3K/AKT pathway in CRC cells transfected with CASC11 overexpression, followed by treatment with LY294002. (B) The main molecules of the PI3K/AKT pathway in CRC cells co-transfected with shCASC11 and the miR-646 or miR-381-3p inhibitor were analyzed. (C) The main molecules of the PI3K/AKT pathway in CRC cells co-transfected with CASC11 overexpression and miR-646 or miR-381-3p mimics were shown. (D, E) IF microscopy of the localization and expression of RAB11FIP2, AKT and p-AKT in CASC11-knockdown cells co-transfected with the miR-646 or miR-381-3p inhibitor.